PXD051741

PXD051741 is an original dataset announced via ProteomeXchange.

Title	KNL2 degradation is regulated by APC/C ubiquitin ligase
Description	Centromeres are specialized chromosomal regions where kinetochore proteins form a protein complex that regulates the cell cycle. KNL2 is an essential protein for kinetochore assembly and plays a significant role in the loading of CENH3. Therefore, the level of KNL2 in plants and other organisms needed to be tightly regulated. Overexpressed AtKNL2 is shown to be degraded via the proteasome degradation pathway by applying an MG115 inhibitor. However, the molecular mechanisms behind the degradation pathway of KNL2 is unknown. Therefore, here we show that the APC/C-dependent degradation of KNL2 is required for maintaining the level and localization of KNL2. We confirmed that the ubiquitin-proteasome system polyubiquitinates and degrades KNL2. Further, IP-MS and Y2H library screening assays performed with KNL2 identified 32 proteins from the ubiquitination pathway. The direct interaction of KNL2 with APC2 was identified using BiFC interaction analysis. Further, the APC10 and Cdc20, core subunit of APC/C, interacts directly with KNL2 as shown by BiFC and Y2H, which is required for substrate recruitment. Moreover, Apcin inhibitor treatment suppresses the degradation of KNL2 in Arabidopsis, validating the involvement of APC/C in KNL2 degradation. Furthermore, our findings highlight the significance of the conserved D-box1 region as a degron for KNL2 in Arabidopsis, facilitating its interaction with APC/C. Further, the interaction of ubiquitin E2 conjugating enzymes (UBC19 and UBC20) with KNL2 has been demonstrated and transfer ubiquitin protein to the lysine sites 336K and 339K of KNL2. Thus, our findings advance our knowledge of cell cycle dynamics by providing insightful information about the molecular mechanisms governing KNL2 regulation.
HostingRepository	PRIDE
AnnounceDate	2025-07-14
AnnouncementXML	Submission_2025-07-13_16:49:57.656.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD051741
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Astrid Bruckmann
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	S-carboxamidoethyl-L-cysteine; acetylated residue; monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	maXis

Revision	Datetime	Status	ChangeLog Entry
0	2024-04-25 08:51:36	ID requested
⏵ 1	2025-07-13 16:49:58	announced

10.6019/PXD051741;

10.1093/plcell/koaf164;

Kalidass M, Jarubula VG, Ratnikava M, Chandra JR, Le Goff S, Probst AV, Esposito S, Grasser KD, Bruckmann A, Gagneux JF, Pros, é, e RF, Rutten T, Schubert V, Demidov D, Lechner E, Steiner FA, Genschik P, Lermontova I, Ubiquitin-dependent proteolysis of KNL2 driven by APC/CCDC20 is critical for centromere integrity and mitotic fidelity. Plant Cell, 37(7):(2025) [pubmed]

submitter keyword: Kinetochore Null2,Centromeres, E3 ligases, E2 conjugating enzymes, Protein-protein interactions, APC/C, D-box

Dr. Astrid Bruckmann
contact affiliation	University Regensburg, Biochemistry I,mass spectrometry group
contact email	astrid.bruckmann@ur.de
lab head
Astrid Bruckmann
contact affiliation	University Regensburg
contact email	astrid.bruckmann@ur.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD051741
     1. Label: PRIDE project
     2. Name: KNL2 degradation is regulated by APC/C ubiquitin ligase